Using near-infrared reflectance spectroscopy (NIRS) to predict the nitrogen levels in the stem and root tissues of Brassica juncea (Indian mustard).

Brassica juncea depends heavily on nitrogen (N) fertilizers for growth and accumulation of seed protein. However, it is an inefficient mobilizer of applied N which leads to accumulation of excess N in the soil, posing environmental risks. Hence, it is imperative to systematically examine spatial-temporal pattern of crop N to efficiently manage N application. The Kjeldahl method is commonly used to estimate N status of crops but it is a destructive method that entails the use of perilous and expensive chemicals. Near-infrared reflectance spectroscopy (NIRS) offers a safe, accurate, and non-destructive alternative for large-scale screening of seed metabolites. Currently, no NIRS model exists to quickly estimate N content in shoots and roots from large germplasm sets in any rapeseed-mustard crop. Developing such a model is essential to breed for enhanced nitrogen use efficiency (NUE). We used 738 shoot and 346 root samples from a B. juncea diversity set to construct the NIRS models. A diverse range of genetic variation in N content was recorded in the stem (0.21-6.61%) and root (0.15-3.04%) tissues of the crop raised on two different N levels (N0 and N100). Modified partial least squares (MPLS) method was employed to establish a regression equation linking reference N values with spectral changes. The developed models exhibited strong associations with reference values, with RSQ values of 0.884 for stem and 0.645 for roots. Furthermore, external validation confirms the reliability of the developed models. The developed models have strong predictive capabilities for rapid and reliable N estimation in various tissues of B. juncea plants.

Isolation and characterization of native antagonistic rhizobacteria against Fusarium wilt of chilli to promote plant growth.

In the eastern coastal regions of Odisha, wilt caused by Fusarium oxysporum f. sp.capsici is an extremely damaging disease in chilli. This disease is very difficult to manage with chemical fungicides since it is soil-borne in nature. The natural rhizosphere soil of the chilli plant was used to isolate and test bacterial antagonists for their effectiveness and ability to promote plant growth. Out of the fifty-five isolates isolated from the rhizosphere of healthy chilli plants, five isolates, namely Iso 01, Iso 17, Iso 23, Iso 24, and Iso 32, showed their highly antagonistic activity against F. oxysporum f. sp. capsici under in vitro. In a dual culture, Iso 32 (73.3%) and Iso 24 (71.5%) caused the highest level of pathogen inhibition. In greenhouse trials, artificially inoculated chilli plants treated with Iso 32 (8.8%) and Iso 24 (10.2%) had decreased percent disease incidence (PDI), with percent disease reduction over control of 85.6% and 83.3%, respectively. Iso 32 and Iso 24 treated chilli seeds have shown higher seed vigor index of 973.7 and 948.8, respectively, as compared to untreated control 636.5. Furthermore, both the isolates significantly increased plant height as well as the fresh and dry weight of chilli plants under the rolled paper towel method. Morphological, biochemical, and molecular characterization identified Bacillus amyloliquefaciens (MH491049) as the key antagonist. This study demonstrates that rhizobacteria, specifically Iso 32 and Iso 24, can effectively protect chilli plants against Fusarium wilt while promoting overall plant development. These findings hold promise for sustainable and eco-friendly management of Fusarium wilt in chilli cultivation.

Rapid identification of reproductive toxicants among environmental chemicals using an in vivo evaluation of gametogenesis in budding yeast Saccharomyces cerevisiae.

Infertility affects ∼12 % of couples, with environmental chemical exposure as a potential contributor. Of the chemicals that are actively manufactured, very few are assessed for reproductive health effects. Rodents are commonly used to evaluate reproductive effects, which is both costly and time consuming. Thus, there is a pressing need for rapid methods to test a broader range of chemicals. Here, we developed a strategy to evaluate large numbers of chemicals for reproductive toxicity via a yeast, S. cerevisiae high-throughput assay to assess gametogenesis as a potential new approach method (NAM). By simultaneously assessing chemicals for growth effects, we can distinguish if a chemical affects gametogenesis only, proliferative growth only or both. We identified a well-known mammalian reproductive toxicant, bisphenol A (BPA) and ranked 19 BPA analogs for reproductive harm. By testing mixtures of BPA and its analogs, we found that BPE and 17 ß-estradiol each together with BPA showed synergistic effects that worsened reproductive outcome. We examined an additional 179 environmental chemicals including phthalates, pesticides, quaternary ammonium compounds and per- and polyfluoroalkyl substances and found 57 with reproductive effects. Many of the chemicals were found to be strong reproductive toxicants that have yet to be tested in mammals. Chemicals having affect before meiosis I division vs. meiosis II division were identified for 16 gametogenesis-specific chemicals. Finally, we demonstrate that in general yeast reproductive toxicity correlates well with published reproductive toxicity in mammals illustrating the promise of this NAM to quickly assess chemicals to prioritize the evaluation for human reproductive harm.

Analyzing high-throughput assay data to advance the rapid screening of environmental chemicals for human reproductive toxicity.

While high-throughput (HTP) assays have been proposed as platforms to rapidly assess reproductive toxicity, there is currently a lack of established assays that specifically address germline development/function and fertility. We assessed the applicability domains of yeast (S. cerevisiae) and nematode (C. elegans) HTP assays in toxicity screening of 124 environmental chemicals, determining their agreement in identifying toxicants and their concordance with reproductive toxicity in vivo. We integrated data generated in the two models and compared results using a streamlined, semi-automated benchmark dose (BMD) modeling approach. We then extracted and modeled relevant mammalian in vivo data available for the matching chemicals included in the Toxicological Reference Database (ToxRefDB). We ranked potencies of common compounds using the BMD and evaluated correlation between the datasets using Pearson and Spearman correlation coefficients. We found moderate to good correlation across the three data sets, with r = 0.48 (95% CI: 0.28-1.00, p<0.001) and rs = 0.40 (p=0.002) for the parametric and rank order correlations between the HTP BMDs; r = 0.95 (95% CI: 0.76-1.00, p=0.0005) and rs = 0.89 (p=0.006) between the yeast assay and ToxRefDB BMDs; and r = 0.81 (95% CI: 0.28-1.00, p=0.014) and rs = 0.75 (p=0.033) between the worm assay and ToxRefDB BMDs. Our findings underscore the potential of these HTP assays to identify environmental chemicals that exhibit reproductive toxicity. Integrating these HTP datasets into mammalian in vivo prediction models using machine learning methods could further enhance the predictive value of these assays in future rapid screening efforts.

Ferromagnetic resonance measurement with frequency modulation down to 2 K.

Ferromagnetic resonance (FMR) spectroscopy is a powerful technique to study the precessional dynamics of magnetization in thin film heterostructures. It provides valuable information about the mechanisms of exchange bias, spin angular momentum transfer across interfaces, and excitation of magnons. A key desirable feature of FMR spectrometers is the capability to study magnetization dynamics over a wide phase space of temperature (T), frequency (f), and magnetic field (B). The design, fabrication, and testing of such a spectrometer, which uses frequency modulation techniques for improved detection of microwave absorption, reduces heat load in the cryostat and allows simultaneous measurements of inverse spin Hall effect (ISHE) induced dc voltages, is described in this paper. The apparatus is based on a 2-port transmitted microwave signal measurement using a grounded co-planar waveguide. The input radio frequency (RF) signal, frequency modulated at a tunable f-band, excites spin precession in the sample, and the attenuated RF signal is measured phase sensitively. The sample stage, inserted in the bore of a superconducting solenoid, allows magnetic field and temperature variability of 0 to ±5 T and 2-310 K, respectively. We demonstrate the working of this Cryo-FMR and ISHE spectrometer on thin films of Ni80Fe20 and Fe60Co20B20 over a wide T, B, and f phase space.

Decennary Update on Oxidative-Rearrangement Involving 1,2-Aryl C-C Migration Around Alkenes: Synthetic and Mechanistic Insights.

In recent years, numerous methodologies on oxidative rearrangements of alkenes have been investigated, that produce multipurpose synthons and heterocyclic scaffolds of potential applications. The present review focused on recently established methodologies for oxidative transformation via 1,2-aryl migration in alkenes (2013-2023). Special emphasis has been placed on mechanistic pathways to understand the reactivity pattern of different substrates, challenges to enhance selectivity, the key role of different reagents, and effect of different substituents, and how they affect the rearrangement process. Moreover, synthetic limitations and future direction also have been discussed. We believe, this review offers new synthetic and mechanistic insight to develop elegant precursors and approaches to explore the utilization of alkene-based compounds for natural product synthesis and functional materials.

Biophysical Mechanisms of Vaginal Smooth Muscle Contraction: The Role of the Membrane Potential and Ion Channels.

The vagina is an essential component of the female reproductive system and is responsible for providing female sexual satisfaction. Vaginal smooth muscle contraction plays a crucial role in various physiological processes, including sexual arousal, childbirth, and urinary continence. In pathophysiological conditions, such as pelvic floor disorders, aberrations in vaginal smooth muscle function can lead to urinary incontinence and pelvic organ prolapse. A set of cellular and sub-cellular physiological mechanisms regulates the contractile properties of the vaginal smooth muscle cells. Calcium influx is a crucial determinant of smooth muscle contraction, facilitated through voltage-dependent calcium channels and calcium release from intracellular stores. Comprehensive reviews on smooth muscle biophysics are relatively scarce within the scientific literature, likely due to the complexity and specialized nature of the topic. The objective of this review is to provide a comprehensive description of alterations in the cellular physiology of vaginal smooth muscle contraction. The benefit associated with this particular approach is that conducting a comprehensive examination of the cellular mechanisms underlying contractile activation will enable the creation of more targeted therapeutic agents to control vaginal contraction disorders.

Genetic diversity, population structure and marker-trait associations in Indian kale (Brassica oleracea L. gp. acephala) using cross-species microsatellite markers.

Kale is known for its exceptional nourishing and functional benefits to human body. However, it is an understudied species from genomic as well as agronomic aspects. It is important to characterize niche kale germplasms around the world to systematically conserve and utilize its genetic variability, especially for commercial traits in the interest of growers, consumers and industry. With this view, genomic and phenotypic characterizations of 62 Kashmiri kale accessions including popular landraces were done to estimate and partition genetic diversity, understand trait relationships, develop population structure and divulge marker-trait associations of economic significance. Sixty-six cross species microsatellite (SSR) markers within Brassica genus amplified 269 alleles in the germplasm. Their polymorphic information content (PIC) ranged from 0.00078 to 0.953 with an average of 0.407. The population structure analysis and neighbour joining tree clustering categorized the germplasm into three sub-populations. AMOVA revealed more within-population variance (67.73 %) than among-populations (32.27 %) variance. The principal component analysis (PCA) involving 24 agronomical traits revealed seven PCs (PC1 to PC7) having Eigen values more than 1, which explained a cumulative variation of 69.21 %. Association mapping with respect to these 24 agronomical traits using mixed linear model and general linear model revealed six overlapping significant marker-trait relationships with five being significant at probability value of 0.001/0.0001. The highly significant associations of two SSRs with economically important traits (siliqua length and seed weight) significantly correlated/related with leaf yield and seed yield were revealed for their possible utilization in marker assisted breeding for higher leaf and seed yields.

Deciphering Winter Sprouting Potential of Erianthus procerus Derived Sugarcane Hybrids under Subtropical Climates.

Winter sprouting potential and red rot resistance are two key parameters for successful sugarcane breeding in the subtropics. However, the cultivated sugarcane hybrids had a narrow genetic base; hence, the present study was planned to evaluate the Erianthus procerus genome introgressed Saccharum hybrids for their ratooning potential under subtropical climates and red rot tolerance under tropical and subtropical climates. A set of 15 Erianthus procerus derived hybrids confirmed through the 5S rDNA marker, along with five check varieties, were evaluated for agro-morphological, quality, and physiological traits for two years (2018-2019 and 2019-2020) and winter sprouting potential for three years (2018-2019, 2019-2020, and 2020-2021). The experimental material was also tested against the most prevalent isolates of the red rot pathogen in tropical (Cf671 and Cf671 + Cf9401) and subtropical regions (Cf08 and Cf09). The E. procerus hybrid GU 12-19 had the highest winter sprouting potential, with a winter sprouting index (WSI) of 10.6, followed by GU 12-22 with a WSI of 8.5. The other top-performing hybrids were as follows: GU 12-21 and GU 12-29 with a WSI of 7.2 and 6.9, respectively. A set of nine E. procerus-derived hybrids, i.e., GU04 (28) EO-2, GU12-19, GU12-21, GU12-22, GU12-23, GU12-26, GU12-27, GU12-30, and GU12-31, were resistant to the most prevalent isolates of red rot in both tropical and subtropical conditions. The association analysis revealed significant correlations between the various traits, particularly the fibre content, with a maximum number of associations, which indicates its multifaceted impact on sugarcane characteristics. Principal component analysis (PCA) summarised the data, explaining 57.6% of the total variation for the measured traits and genotypes, providing valuable insights into the performance and characteristics of the Erianthus procerus derived hybrids under subtropical climates. The anthocyanin content of Erianthus procerus hybrids was better than the check varieties, ranging from 0.123 to 0.179 (2018-2019) and 0.111 to 0.172 (2019-2020); anthocyanin plays a vital role in mitigating cold injury, acting as an antioxidant in cool weather conditions, particularly in sugarcane. Seven hybrids recorded a more than 22% fibre threshold, indicating their industrial potential. These hybrids could serve as potential donors for cold tolerance and a high ratooning ability, along with red rot resistance, under subtropical climates.

Mitigating postharvest quantitative and qualitative losses in mango fruits through the application of biocontrol agents: An in-vivo and in-vitro assessment.

Mango is a commercial fruit crop of India that suffers huge postharvest losses every year. The application of biocontrol agents (BCAs) bears a vast potential for managing the same, which is yet to be exploited to its fullest extent. Hence, studies were conducted for BCAs application of Debaryomyces hansenii, Bacillus subtilis and Pseudomonas fluorescens strains on mango fruit under in-vitro, in-vivo conditions to know the efficacy of these BCAs on the postharvest pathogen, shelf life and quality retention of mango fruit. The 'poisoned food technique' was attempted for in-vitro studies. For the in-vivo studies, fruit of the commercial cultivar 'Amrapali' were un-inoculated and pre-inoculated with major postharvest pathogens (anthracnose: Colletotrichum gloeosporioides and stem-end rot: Botryodiplodia theobromae) were treated with BCA, followed by ambient storage at (24 ± 4 °C, 75 ± 5 % RH). From the results, it has been observed that under in vitro studies, BCA Debaryomyces hansenii (Strain: KP006) and Bacillus subtilis (Strain: BJ0011) at the treatment level 108 CFU mL-1 while, the Pseudomonas fluorescens at 109 CFU mL-1 (Strain: BE0001) were significantly effective for pathogen inhibition. However, under the in vivo studies, the BCA Debaryomyces hansenii (Strain: KP006) at 108 CFU mL-1 treatment level was found to significantly reduce the pathogen's decay incidence while positively influencing the shelf life and biochemical (quality) attributes. This treatment increased the storage life of mango fruit by more than three days over control fruit. Therefore, BCA Debaryomyces hansenii (Strain: KP006) at 108 CFU mL-1 can be used to control the postharvest pathological loss of mango fruit without affecting its internal quality.

Biomass and energy potential of Erianthus arundinaceus and Saccharum spontaneum-derived novel sugarcane hybrids in rainfed environments.

BACKGROUND: Energy canes are viable feedstocks for biomass industries due to their high biomass production potential, lower susceptibility to insects and diseases, better ability to adapt to extreme conditions and clean bioenergy. Interspecific hybrids (ISH) and intergeneric hybrids (IGH) have great potential to meet the growing demand of biomass, biomass-derived energy and feedstock. RESULTS: In this study, two types of energy canes, Type I and Type II, derived from S. spontaneum and E. arundinaceous background were evaluated for high biomass, fiber and bioenergy potential under subtropical climate along with the check varieties Co 0238 and CoS 767. Out of 18 energy canes studied, six energy canes, viz., SBIEC11008 (204.15 t/ha), SBIEC11005 (192.93 t/ha), SBIEC13008 (201.26 t/ha), SBIEC13009 (196.58 t/ha), SBIEC13002 (170.15 t/ha), and SBIEC13007 (173.76 t/ha), consistently outperformed the check varieties under Type-I, whereas in type-II, SBIEC11004 (225.78 t/ha), SBIEC11006 (184.89 t/ha), and SBIEC14006 (184.73 t/ha) energy canes produced significantly higher biomass than commercial checks, indicating their superior potential for cogeneration. Estimated energy output from the energy canes (700-1300 GJ/ha/year) exceeded the range of co-varieties (400-500 GJ/ha/year) and energy utilization efficiency in plants and ratoon crops for energy canes viz., SBIEC11008 (3%, 1.97%), SBIEC14006 (1.93%, 2.4%), SBIEC11005 (1.7%, 1.9%), and SBIEC11001 (1.01%, 1.03%), was higher than best checks Co 0238 (0.77, 0.9%). Additionally, energy canes SBIEC 13001 (22.35%), SBIEC 11008 (22.50%), SBIEC 14006 (28.54%), SBIEC 11004 (30.17%) and SBIEC 11001 (27.03%) had higher fiber contents than the co-varieties (12.45%). CONCLUSION: The study gives insight about the potential energy canes for higher biomass and energy value. These energy cane presents a vital option to meet the future demand of bioenergy, fiber and fodder for biomass due to their versatile capacity to grow easily under marginal lands without competing with cultivated land worldwide.

Salt stress influences the proliferation of Fusarium solani and enhances the severity of wilt disease in potato.

Soil salinity has emerged as a critical abiotic stress in potato production, whereas wilt disease, caused by Fusarium solani, is the significant biotic stress. An experiment was performed to decipher the occurrence of wilt incidence by F. solani FJ1 under the influence of salinity in both in vitroand pot culture conditions. High salt concentration negatively influenced root and shoot development in the variety "Kufri Jyoti" but positively affected the mycelial growth and sporulation behaviours of F. solani FJ1. There was abundant whitish mycelial growth with enhanced biomass and high sporulation (microconidia production) in F. solani FJ1 cultured on salt-supplemented media. Moreover, under high salinity conditions (EC 2-8 dS m-1), severe wilting and rotting of vascular bundles were observed in plants artificially inoculated with F. solani FJ1. The mortality rate of potato plants was significantly higher under individual and combined stresses as compared to control. The wilt index of individual and combined stressed plants was also substantially higher compared to the control. Additionally, compared to the control, there was a significant decrease in total chlorophyll content and membrane stability index of the leaves under combined stress. However, the total phenols were increased under stress conditions. The total sugar content of potato plants decreased in infected plants, but increased when exposed to salt stress or a combination of salt stress and pathogen infection. F. solani infection also increased the activity of peroxidase (POX) and decreased the activity of phenylalanine ammonia-lyase (PAL) and catalase (CAT). These results suggest that Fusarium wilt and dry rot will be a more severe disease for potato cultivation in saline soils.

Yeast-Based Screening of Anti-Viral Molecules.

Viruses are minuscule infectious agents that reproduce exclusively within the living cells of an organism and are present in almost every ecosystem. Their continuous interaction with humans poses a significant threat to the survival and well-being of everyone. Apart from the common cold or seasonal influenza, viruses are also responsible for several important diseases such as polio, rabies, smallpox, and most recently COVID-19. Besides the loss of life and long-term health-related issues, clinical viral infections have significant economic and social impacts. Viral enzymes, especially proteases which are essential for viral multiplication, represent attractive drug targets. As a result, screening of viral protease inhibitors has gained a lot of interest in the development of anti-viral drugs. Despite the availability of anti-viral therapeutics, there is a clear need to develop novel curative agents that can be used against a given virus or group of related viruses. This review highlights the importance of yeasts as an in vivo model for screening viral enzyme inhibitors. We also discuss the advantages of yeast-based screening platforms over traditional assays. Therefore, in the present article, we discuss why yeast is emerging as a model of choice for in vivo screening of anti-viral molecules and why yeast-based screening will become more relevant in the future for screening anti-viral and other molecules of clinical importance.

New Trends in Vaccine Characterization, Formulations, and Development.

More than eight decades have passed since the development of the first vaccine in the 1940s [...].

Glycogen Granules Are Degraded by Non-Selective Autophagy in Nitrogen-Starved Komagataella phaffii.

Autophagy was initially recognized as a bulk degradation process that randomly sequesters and degrades cytoplasmic material in lysosomes (vacuoles in yeast). In recent years, various types of selective autophagy have been discovered. Glycophagy, the selective autophagy of glycogen granules, is one of them. While autophagy of glycogen is an important contributor to Pompe disease, which is characterized by the lysosomal accumulation of glycogen, its selectivity is still a matter of debate. Here, we developed the Komagataella phaffii yeast as a simple model of glycogen autophagy under nitrogen starvation conditions to address the question of its selectivity. For this, we turned the self-glucosylating initiator of glycogen synthesis, Glg1, which is covalently bound to glycogen, into the Glg1-GFP autophagic reporter. Our results revealed that vacuolar delivery of Glg1-GFP and its processing to free GFP were strictly dependent on autophagic machinery and vacuolar proteolysis. Notably, this process was independent of Atg11, the scaffold protein common for many selective autophagy pathways. Importantly, the non-mutated Glg1-GFP (which synthesizes and marks glycogen) and mutated Glg1Y212F-GFP (which does not synthesize glycogen and is degraded by non-selective autophagy as cytosolic Pgk1-GFP) were equally well delivered to the vacuole and had similar levels of released GFP. Therefore, we concluded that glycogen autophagy is a non-selective process in K. phaffii yeast under nitrogen starvation conditions.

An investigation of the environmental implications of bioplastics: Recent advancements on the development of environmentally friendly bioplastics solutions.

The production and utilization of plastics may prove beneficial, but the environmental impact suggests the opposite. The single-use plastics (SUP) and conventional plastics are harmful to the environment and need prompt disposal. Bioplastics are increasingly being considered as a viable alternative to conventional plastics due to their potential to alleviate environmental concerns such as greenhouse gas emissions and pollution. However, the previous reviews revealed a lack of consistency in the methodologies used in the Life Cycle Assessments (LCAs), making it difficult to compare the results across studies. The current study provides a systematic review of LCAs that assess the environmental impact of bioplastics. The different mechanical characteristics of bio plastics, like tensile strength, Young's modulus, flexural modulus, and elongation at break are reviewed which suggest that bio plastics are comparatively much better than synthetic plastics. Bioplastics have more efficient mechanical properties compared to synthetic plastics which signifies that bioplastics are more sustainable and reliable than synthetic plastics. The key challenges in bioplastic adoption and production include competition with food production for feedstock, high production costs, uncertainty in end-of-life management, limited biodegradability, lack of standardization, and technical performance limitations. Addressing these challenges requires collaboration among stakeholders to drive innovation, reduce costs, improve end-of-life management, and promote awareness and education. Overall, the study suggests that while bioplastics have the potential to reduce environmental impact, further research is needed to better understand their life cycle and optimize their end-of-life (EoL) management and production to maximize their environmental benefits.

Transition-Metal-Catalyzed 1,2-Diaminations of Olefins: Synthetic Methodologies and Mechanistic Studies.

1,2-Diamines are synthetically important motifs in organo-catalysis, natural products, and drug research. Continuous utilization of transition-metal based catalyst in direct 1,2-diamination of olefines, in contrast to metal-free transformations, with numerous impressive advances made in recent years (2015-2023). This review summarized contemporary research on the transition-metal catalyzed/mediated [e. g., Cu(II), Pd(II), Fe(II), Rh(III), Ir(III), and Co(II)] 1,2-diamination (asymmetric and non-asymmetric) especially emphasizing the recent synthetic methodologies and mechanistic understandings. Moreover, up-to-date discussion on (i) paramount role of oxidant and catalyst (ii) key achievements (iii) generality and uniqueness, (iv) synthetic limitations or future challenges, and (v) future opportunities are summarized related to this potential area.

Imidazo[1,2-a]pyrimidine as a New Antileishmanial Pharmacophore against Leishmania amazonensis Promastigotes and Amastigotes.

Leishmania poses a substantial threat to the human population all over the globe because of its visceral and cutaneous spread engendered by all 20 species. Unfortunately, the available drugs against leishmania are already hobbled with toxicity, prolonged treatment, and increasing instances of acquirement of resistance. Under these grave circumstances, the development of new drugs has become imperative to keep these harmful microbes at bay. To this end, a Groebke-Blackburn-Bienaymé multicomponent reaction-based library of different imidazo-fused heterocycles has been synthesized and screened against Leishmania amazonensis promastigotes and amastigotes. Among the library compounds, the imidazo-pyrimidine 24 has been found to be the most effective (inhibitory concentration of 50% (IC50) < 10 µM), with selective antileishmanial activity on amastigote forms, a stage of the parasite related to human disease. The compound 24 has exhibited an IC50 value of 6.63 µM, being ∼two times more active than miltefosine, a reference drug. Furthermore, this compound is >10 times more destructive to the intracellular parasites than host cells. The observed in vitro antileishmanial activity along with suitable in silico physicochemical and absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties of compound 24 reinforce the imidazo-pyrimidine scaffold as a new antileishmanial pharmacophore and encourage further murine experimental leishmaniasis studies.

Imidazo[2,1-b]thiazole based indoleamine-2,3-dioxygenase 1 (IDO1) inhibitor: Structure based design, synthesis, bio-evaluation and docking studies.

Indoleamine-2,3-dioxygenase 1 (IDO1) is an immunomodulatory enzyme known to catalyse the initial and rate limiting step of kynurenine pathway of l-tryptophan metabolism. IDO1 enzyme over expression plays a crucial role in progression of cancer, malaria, multiple sclerosis and other life-threatening diseases. Several efforts over the last two decades have been invested by the researchers for the discovery of different IDO1 inhibitors and the plasticity of the IDO1 enzyme ligand binding pocket provide ample opportunities to develop new heterocyclic scaffolds targeting this enzyme. In the present work, based on the X-ray crystal structure of human IDO1 coordinated with few ligands, we designed and synthesized new fused heterocyclic compounds and evaluated their potential human IDO1 inhibitory activity (compound 30 and 41 showed IC50 values of 23 and 13 µM, respectively). The identified HITs were observed to be non-toxic to HEK293 cells at 100 µM concentration. The observed activity of the synthesized compounds was correlated with the specific interactions of their structures at the enzyme pocket using docking studies. A detailed analysis of docking results of the synthesized analogues as well as selected known IDO1 inhibitors revealed that most of the inhibitors have some reasonable docking scores in at least two crystal structures and have similar orientation as that of co-crystal ligands.

Optimization of a simple, low-cost one-step reverse transcription recombinase polymerase amplification method for real-time detection of potato virus A in potato leaves and tubers.

Vegetative propagation of potatoes makes it possible for potato viruses to be transmitted through tubers. Potato virus A (PVA) is one of these viruses, which belongs to the Potyvirus genus in the Potyviridae family. Potato tuber yield can be reduced by 30-40% by PVA alone. Losses can be further exacerbated by potato virus X and/or potato virus Y infection. PVA is transmitted primarily by several species of aphids in non-persistent manner. With the aim of resolving this problem, we developed one-step reverse transcription-recombinase polymerase amplification (RT-RPA), a highly sensitive and cost-effective method for detecting PVA in both potato tubers and leaves. Detection and amplification are performed using isothermal conditions in this method. There was good amplification of the coat protein gene in PVA with all three primers tested. To conduct this study, a primer set that can amplify specific 185 base pair (bp) product was selected. PVA detection was optimized by 30-min amplification reactions, which showed no cross-reactivity with other potato viruses. A simple heating block or water bath was used to amplify PVA product using RT-RPA at a temperature range of 38-42 °C. In comparison to conventional reverse transcription-polymerase chain reaction (RT-PCR), the newly developed RT-RPA protocol exhibited high sensitivity for both potato leaves and tuber tissues. Using cellular paper-based simple RNA extraction procedure, the virus was detected in leaf samples as efficiently as purified total RNA. We also found that combining LiCl-based RNA precipitation with cellular paper discs allowed us to successfully optimize RNA extraction for one-step RT-RPA for detecting PVA in tubers. Tests using this simplified one-step RT-RPA method were successfully applied to 300 samples of both leaves and tubers from various potato cultivars. In our knowledge, this is the first report of an RT-RPA assay utilizing simple RNA obtained from either cellular disc paper or LiCl coupled with cellular disc paper to detect PVA. As a result, this method was equally sensitive and specific for detecting PVA in potatoes. The developed RT-RPA assay is more versatile, durable, and do not require highly purified RNA templates, thus providing an effective alternative to RT-PCR assays for screening of germplasm, certifying planting materials, breeding for virus resistance, and real-time monitoring of PVA.